Coding Sequence Validation

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The Mouse Biology Program offers Coding Sequence Validation as part of our comprehensive genetic analysis services. This process is critical for confirming that the coding region of a targeted gene has been correctly modified, ensuring precise genetic alterations in mouse models. Coding sequence validation is essential for maintaining the integrity and functionality of engineered models, whether for knockouts, knock-ins, or other gene modifications.

What is Coding Sequence Validation?

Coding sequence validation involves thoroughly examining the modified coding region within a gene to ensure that the desired changes have been accurately introduced. This process helps confirm that the sequence is free from unintended mutations and maintains proper reading frames, which is crucial for generating functional genetic models.

How Coding Sequence Validation Works

• Sanger Sequencing:
  The primary method for coding sequence validation is Sanger sequencing, which provides a detailed, base-by-base confirmation of the modified gene. This approach ensures that the correct nucleotide changes have been made and the reading frame remains intact.
• RT-PCR Analysis:
  For specific projects, reverse transcription PCR (RT-PCR) may be used to confirm that the modified coding sequence is transcribed adequately into mRNA. This step helps ensure that the gene remains functional after modification.
• In Silico Analysis:
  Bioinformatics tools analyze sequencing data, verifying that the edited coding sequence aligns with the intended genetic design. This step detects potential off-target effects or unintended mutations, providing comprehensive validation.

Why is Coding Sequence Validation Important?

• Ensuring Functional Integrity:
  It confirms that the genetic modification maintains the correct reading frame and produces the desired protein, supporting valid experimental outcomes.
• Preventing Off-Target Effects:
  Validation helps detect and correct unintended mutations, reducing the risk of confounding results in downstream research.
• Maintaining Research Accuracy:
  Reliable coding sequence validation ensures that only accurately modified models are used, leading to consistent and reproducible study results.

Precision and Reliability in Genetic Modifications
Our coding sequence validation service ensures that your genetic modifications are accurate, functional, and ready to support meaningful research outcomes.

How to Request Coding Sequence Validation

Please contact our team to request coding sequence validation. We will provide detailed guidance on sample submission, service timelines, and data analysis to ensure your gene modification project meets the highest standards.

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• Genotyping Services
• Coding Sequence Validation
• Genetic Analysis Services
• Genomic Insertion Site Identification
• Homologous Recombination Confirmation in ES Cells
• Off-Target Mutagenesis Detection